Paternity determination of interspecific rhododendron hybrids by genomic in situ hybridization (GISH)

Genome ◽  
2010 ◽  
Vol 53 (4) ◽  
pp. 277-284 ◽  
Author(s):  
Małgorzata Czernicka ◽  
Anna Mścichowska ◽  
Maria Klein ◽  
Piotr Muras ◽  
Ewa Grzebelus
Keyword(s):  
In Situ Hybridization ◽  
Mitotic Chromosome ◽  
Genomic In Situ Hybridization ◽  
Interspecific Crosses ◽  
Practical Tool ◽  
Paternity Determination ◽  
Hybrid Character ◽  
Positive Results

Genomic in situ hybridization (GISH) has been proved to be the most effective and accurate technique for confirmation of hybrid character. The objective of our study was to adapt and optimize a GISH protocol for identification of donor chromatin in hybrids obtained by interspecific crosses between five Rhododendron taxa ( R. aureum , R. brachycarpum , R. catawbiense ‘Catharine van Tol’, R. catawbiense ‘Nova Zembla’, and R. yakushimanum ‘Koichiro Wada’). Positive results were obtained only when we used mitotic chromosome spreads prepared from anthers. The best differentiation of maternal and paternal chromosomes in hybrid genomes was obtained when 50 ng of probe was applied together with blocking DNA at a concentration of 3.0 µg/µL. The results demonstrate that GISH is a practical tool for detection of alien genomes and analysis of the constitution of the chromosomes in rhododendron hybrids.

Characterization by RFLP analysis and genomic in situ hybridization of a recombinant and a monosomic substitution plant derived from Hordeum vulgare L. × Hordeum bulbosum L. crosses

Genome ◽  
1997 ◽  
Vol 40 (2) ◽  
pp. 195-200 ◽  
Author(s):  
R. A. Pickering ◽  
A. M. Hill ◽  
R. G. Kynast
Keyword(s):  
In Situ Hybridization ◽  
Hordeum Vulgare ◽  
Rflp Analysis ◽  
Leaf Sheath ◽  
Genomic In Situ Hybridization ◽  
Barley Chromosome ◽  
Interspecific Crosses ◽  
Hordeum Bulbosum ◽  
Hordeum Vulgare L

Interspecific crosses in Hordeum have been made with the aim of transferring desirable traits, such as disease resistance, from a wild species, Hordeum bulbosum, into cultivated barley (Hordeum vulgare). Interspecific recombinants have previously been identified using several methods, but there are limitations with all the techniques. We improved our ability to characterize progeny from H. vulgare × H. bulbosum crosses by using genomic in situ hybridization (GISH). The plant material comprised a recombinant and a monosomic alien substitution plant derived from H. vulgare × H. bulbosum crosses. The recombinant possesses a pubescent leaf sheath conferred by a gene transferred from H. bulbosum into barley cultivar Golden Promise. The use of GISH on a plant homozygous for the pubescence gene confirmed the presence of H. bulbosum DNA located distally on two barley chromosomes and we mapped the introgression to barley chromosome 4HL using RFLP analysis. Furthermore, by means of an allelism test we found that the transferred gene for pubescence is allelic or closely linked to a gene for pubescence (Hs) located on barley chromosome 4HL. The presence of a single H. bulbosum chromosome in the monosomic substitution plant was confirmed by GISH. A distal introgression of H. bulbosum DNA was also observed on one barley chromosome, which was located on chromosome 3HL by RFLP analysis.Key words: Hordeum vulgare, Hordeum bulbosum, interspecific hybrid, gene introgression, genomic in situ hybridization.

Determination of the Paternity of Wheat ( L) × Jointed Goatgrass ( Host) BC Plants by Using Genomic In Situ Hybridization (GISH) Technique

Crop Science ◽  
2002 ◽  
Vol 42 (3) ◽  
pp. 939 ◽  
Author(s):  
Zhining Wang ◽  
Robert S. Zemetra ◽  
Jennifer Hansen ◽  
An Hang ◽  
Carol A. Mallory-Smith ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Genomic In Situ Hybridization ◽  
Jointed Goatgrass

Genome discrimination in progeny of interspecific hybrids between Brassica napus and Raphanus raphanistrum

Genome ◽  
2003 ◽  
Vol 46 (3) ◽  
pp. 469-472 ◽  
Author(s):  
A Benabdelmouna ◽  
G Guéritaine ◽  
M Abirached-Darmency ◽  
H Darmency
Keyword(s):  
In Situ Hybridization ◽  
Brassica Napus ◽  
Oilseed Rape ◽  
Interspecific Hybrid ◽  
Interspecific Hybrids ◽  
Genomic In Situ Hybridization ◽  
Interspecific Crosses ◽  
Wild Radish ◽  
Raphanus Raphanistrum

Genomic in situ hybridization (GISH) applied to the F1 interspecific hybrid between oilseed rape (Brassica napus, AACC, 2n = 38) and wild radish (Raphanus raphanistrum, RrRr, 2n = 18) showed the predicted 19 chromosomes from B. napus and 9 chromosomes from R. raphanistrum. The very low female fertility of these interspecific hybrids when backcrossed to R. raphanistrum led to only two descendants. Their chromosome number varied between 45 and 48. Both of these progenies showed only 9 chromosomes from R. raphanistrum and 36–39 chromosomes from B. napus. These results indicate the efficiency and limits of GISH as a suitable tool to assess and interpret the behavior of chromosomes after such interspecific crosses. The unexpected chromosome combination is discussed.Key words: genomic in situ hybridization, interspecific hybrid, introgression, oilseed rape, wild radish.

Synthesis and cytological characterization of trigeneric hybrids of durum wheat with and without Ph1

Genome ◽  
2004 ◽  
Vol 47 (6) ◽  
pp. 1173-1181 ◽  
Author(s):  
Prem P Jauhar ◽  
M Doğramaci ◽  
T S Peterson
Keyword(s):  
In Situ Hybridization ◽  
Chromosome Pairing ◽  
Genetic Recombination ◽  
Genomic In Situ Hybridization ◽  
Homoeologous Pairing ◽  
Alien Gene Transfer ◽  
Chromosome 5B ◽  
Alien Gene ◽  
Trigeneric Hybrids

Wild grasses in the tribe Triticeae, some in the primary or secondary gene pool of wheat, are excellent reservoirs of genes for superior agronomic traits, including resistance to various diseases. Thus, the diploid wheatgrasses Thinopyrum bessarabicum (Savul. and Rayss) Á. Löve (2n = 2x = 14; JJ genome) and Lophopyrum elongatum (Host) Á. Löve (2n = 2x = 14; EE genome) are important sources of genes for disease resistance, e.g., Fusarium head blight resistance that may be transferred to wheat. By crossing fertile amphidiploids (2n = 4x = 28; JJEE) developed from F1 hybrids of the 2 diploid species with appropriate genetic stocks of durum wheat, we synthesized trigeneric hybrids (2n = 4x = 28; ABJE) incorporating both the J and E genomes of the grass species with the durum genomes A and B. Trigeneric hybrids with and without the homoeologous-pairing suppressor gene, Ph1, were produced. In the absence of Ph1, the chances of genetic recombination between chromosomes of the 2 useful grass genomes (JE) and those of the durum genomes (AB) would be enhanced. Meiotic chromosome pairing was studied using both conventional staining and fluorescent genomic in situ hybridization (fl-GISH). As expected, the Ph1-intergeneric hybrids showed low chromosome pairing (23.86% of the complement), whereas the trigenerics with ph1b (49.49%) and those with their chromosome 5B replaced by 5D (49.09%) showed much higher pairing. The absence of Ph1 allowed pairing and, hence, genetic recombination between homoeologous chromosomes. Fl-GISH analysis afforded an excellent tool for studying the specificity of chromosome pairing: wheat with grass, wheat with wheat, or grass with grass. In the trigeneric hybrids that lacked chromosome 5B, and hence lacked the Ph1 gene, the wheat–grass pairing was elevated, i.e., 2.6 chiasmata per cell, a welcome feature from the breeding standpoint. Using Langdon 5D(5B) disomic substitution for making trigeneric hybrids should promote homoeologous pairing between durum and grass chromosomes and hence accelerate alien gene transfer into the durum genomes.Key words: alien gene transfer, chiasma (xma) frequency, chromosome pairing, fluorescent genomic in situ hybridization (fl-GISH), homoeologous-pairing regulator, specificity of chromosome pairing, wheatgrass.

Genome constitutions ofRoegneriaalashanica,R.elytrigioides,R.magnicaespesandR.grandis(Poaceae: Triticeae) via genomic in-situ hybridization

2010 ◽  
Vol 28 (2) ◽  
pp. 206-211 ◽  
Author(s):  
Hai-Qing Yu ◽  
Chun Zhang ◽  
Chun-Bang Ding ◽  
Hai-Qin Zhang ◽  
Yong-Hong Zhou
Keyword(s):  
In Situ Hybridization ◽  
Genomic In Situ Hybridization
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